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Virus replication cycle of white spot syndrome virus in secondary cell cultures from the lymphoid organ of Litopenaeus vannamei

Journal Contribution - Journal Article

The replication cycle of white spot syndrome virus (WSSV) was investigated in secondary cell cultures from the lymphoid organ of Litopenaeus vannamei. The secondary cells formed a confluent monolayer at 24 h post-reseeding, and this nnonolayer could be maintained for 10 days with a viability of 90 %. Binding of WSSV to cells reached a maximum (73 +/- 3 % of cells and 4.84 +/- 0.2 virus particles per virus-binding cell) at 120 min at 4 degrees C. WSSV entered cells by endocytosis. The co-localization of WSSV and early endosonnes was observed starting from 30 min post-inoculation (p.i.). Double indirect immunofluorescence staining showed that all cell-bound WSSV particles entered these cells in the period between 0 and 60 min p.i. and that the uncoating of WSSV occurred in the same period. After 1 h inoculation at 27 degrees C, the WSSV nucleocapsid protein VP664 and envelope protein VP28 started to be synthesized in the cytoplasm from 1 and 3 h p.i., and were transported into nuclei from 3 and 6 h p.i., respectively. The percentage of cells that were VP664- and VP28-positive in their nuclei peaked (50 +/- 4 %) at 12 h p.i. Quantitative PCR showed that WSSV DNA started to be synthesized from 6 h p.i. In vivo titration of the supernatants showed that the progeny WSSV were released from 12 h p.i. and peaked at 18 h p.i. In conclusion, the secondary cell cultures from the lymphoid organ were proven to be ideal for examination of the replication cycle of WSSV.
Journal: JOURNAL OF GENERAL VIROLOGY
ISSN: 0022-1317
Issue: 9
Volume: 96
Pages: 2844 - 2854
Publication year:2015
BOF-keylabel:yes
IOF-keylabel:yes
BOF-publication weight:0.1
CSS-citation score:1
Authors:National
Authors from:Higher Education
Accessibility:Closed